1. Adapting Image J to perform Bone Histomorphometry
Goal: Develop method by which Bone histomorphometry can be performed without cost by use of free software from the NIH.

2. Commercial software is used to perform the measurements
Bone histomorphometry uses stained bone slides to measure various bone parameters
Commercial software is used to perform the measurements
But these are all based on basic measurements of length, area, and number
Software specifically tailored towards bone field simplifies the measurements, but little competition leads to high prices
Image J can perform the basic same measurements (Area, length and number)
Length
Area

3. Image J calculates area accurately
Compared Image J calculated vs manually calculated measurements of area from various samples.
Control sample was 2 * 2 square inch box
Started creating void areas within the box to make sure that image J can measure complicated images

5. Image J introduces error with distance measurements of circles
On a computer screen, circles are made of square pixels
Comparing to manual calculations, image J introduced an average error of 5.4%

6. Correcting for error introduced
Error was consistent in that Image J was over estimating the distance and a little over 5%
Excel equation can automatically remove the error introduced by Image J.
Average error was reduced from 5.4% to .29%

7. Next steps
All bone histomorphometry is performed on undecalcified bone samples embedded in methyl methacrylate
We have some bone samples, we need to get the materials to do the embedding
Also need to figure out proper cutting angles for the polycut microtome
Analysis is performed on samples stained using standard H/E and Goldners Trichrome
H/E allows for measurement of bone and tissue volumes
Goldner trichrome reveals unmineralized bone matrix, allowing measurements of osteoid volumes